Characterization of the Taeniaspp HDP2 sequence and development of a novel PCR-based assay for discrimination of Taenia saginatafrom Taenia asiatica

González, Bailo Begoña, Ferrer Elizabeth, García María, Harrison, Parkhouse, Mcmanus, Gárate, Gárate Teresa

Description

A previously described Taenia saginata HDP2 DNA sequence, a 4-kb polymorphic fragment, was previously used as the basis for developing PCR diagnostic protocols for the species-specific discrimination of T. saginata from T. solium and for the differentiation of T. saginata from T. asiatica . The latter was shown subsequently to lack the required specificity, so we undertook genetic studies of the HDP2 sequence from T. saginata and T. asiatica to determine why, and to develop a novel HDP2-PCR protocol for the simultaneous unambiguous identification of human taeniids. Sequencing and further analysis of the HDP2 DNA fragments of 19 Asiatic isolates of T. saginata and T. asiatica indicated that the HDP2 sequences of both species exhibited clear genomic variability, due to polymorphic variable fragments, that could correspond to the non-transcribed region of ribosomal DNA. This newly observed polymorphism allowed us to develop a novel, reproducible and reliable HDP2-PCR protocol which permitted the simultaneous discrimination of all T. saginata and T. asiatica isolates examined. This species-specific identification was based on, and facilitated by, the clear size difference in amplicon profiles generated: fragments of 1300 bp, 600 bp and 300 bp were produced for T. asiatica , amplicons of 1300 bp and 300 bp being obtained for T. saginata . Control T. solium samples produced one amplicon of 600 bp with the HDP2-PCR protocol. The assay has the potential to prove useful as a diagnostic tool in areas such as South East Asia where T. saginata , T. asiatica and T. solium coexist.

Informations

• Publié par: biomed
• Publié le: 01 janvier 2010
• Nombre de lectures: 5
• Langue: English

Extrait

Gonzálezet al.Parasites & Vectors2010,3:51 http://www.parasitesandvectors.com/content/3/1/51

R E S E A R C H Open Access Research Characterization of the Taenia spp HDP2 sequence and development of a novel PCR-based assay for discrimination of Taenia saginata from Taenia asiatica

1 1 2 1 3 Luis M González , Begoña Bailo , Elizabeth Ferrer , Maria D Fernandez García , Leslie JS Harrison , 4 5 1 Michael RE Parkhouse , Donald P McManus and Teresa Gárate*

Findings Human taeniasis results from intestinal infection with the adult tapewormsTaenia saginata,Taenia asiaticaorTae- nia solium. Ingestion of eggs ofT. saginataandT. solium/ T. asiaticacauses cysticercosis in cattle and pigs, respec- tively. Importantly,T. soliumeggs can also infect man, giving rise to human cysticercosis, with the frequent localization of the metacestodes in the brain, causing potentially fatal neurocysticercosis (NCC) [1-3]. Infec- tion withT. soliumis therefore a serious public health problem, notably in endemic areas (Latin America, Africa, Asia), but also in non-endemic areas due to imported cases [4]. Hence, and as an essential part of

* Correspondence: [e-mail] 1 Parasitology Department, Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain Full list of author information is available at the end of the article

transmission control programs, there is an immediate and practical need for reliable and species-specific diag- nosis of human taeniasis caused byT. saginata,T. asiatica andT. soliumin order to identify tapeworm carriers, par- ticularly those withT. solium, thus helping to avoid trans- mission of cysticercosis/NCC. Until recently, species-specific diagnosis of taeniasis was unsatisfactory. Conventional clinical and morpholog- ical identification has low specificity and sensitivity [5]. Moreover, adult worms ofT. saginataandT. asiaticaare frequently confused due to their morphological similari- ties [6]. Copro-antigen diagnosis by ELISA, that detects parasite antigens in patient feces, is based on the use of polyclonal antibody reagents although it does not distin- guish among the human intestinal taeniids [7,8]. More recently, serological diagnosis forT. soliumtaeniasis has

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